Composition and method to improve blood lipid profiles and optionally reduce low density lipoprotein (ldl) per-oxidation in humans

ABSTRACT

A composition and method which improves blood lipid profiles and optionally reduces low density lipoprotein (LDL) per-oxidation in humans by administering a therapeutic amount of a composition comprising krill oil in combination with astaxanthin or a mixture of fish oil derived, choline based, phospholipid bound omega-3 fatty acid mixture including phospholipid bound polyunsaturated EPA and DHA. In one embodiment, the krill oil is derived from  Euphasia  spp., comprising Eicosapentaenoic (EPA) and Docosahexaenoic (DHA) fatty acids in the form of triacylglycerides and phospholipids. The krill oil includes at least 10% EPA and 5% DHA, of which greater than 50% are in the form of phospholipids and the 1-4000 mg of krill oil per daily dose is delivered.

RELATED APPLICATION(S)

This application is based upon prior filed provisional application Ser.No. 61/329,744, filed Apr. 30, 2010; and related to prior filed U.S.patent application Ser. No. 12/840,372, filed Jul. 21, 2010, and Ser.No. 13/079,238, filed Apr. 4, 2011, the disclosures which are herebyincorporated by reference in their entirety.

FIELD OF THE INVENTION

This invention relates to improving blood lipid profiles and reducinglow density lipoprotein (LDL) oxidation using therapeutic compositionsand methods derived from krill oil extracts and/or marine oilcompositions. This invention also relates to improving blood lipidprofiles and reducing LDL using therapeutic composition and methodsderived from a mixture of fish oil and fish oil derived, choline based,phospholipid bound fatty acids including polyunsaturated EPA and DHAeither alone or mixed with synergistic additives.

BACKGROUND OF THE INVENTION

The use of krill and/or marine oil are disclosed in U.S. PatentPublication Nos. 2004/0234587; 2004/0241249; and 2007/0098808, thedisclosures which are hereby incorporated by reference in theirentirety, and discussed in related U.S. patent application Ser. Nos.12/840,372 and 13/079,238. The beneficial aspects of using krill and/ormarine oil are shown also in a research paper published by L. Deutsch as“Evaluation of the Effect of Neptune Krill Oil on Chronic Inflammationand Arthritic Symptoms,” published in the Journal of the AmericanCollege of Nutrition, Volume 26, No. 1, 39-49 (2007), the disclosurewhich is hereby incorporated by reference in its entirety.

The published '587, '249 and '808 applications discuss the beneficialaspects of using krill oil in association with pharmaceuticallyacceptable carriers. As an example, this krill and/or marine oil can beobtained by the combination of detailed steps as taught in the '808application, by placing krill and/or marine material in a ketonesolvent, separating the liquid and solid contents, recovering a firstlipid rich fraction from the liquid contents by evaporation, placing thesolid contents and organic solvent in an organic solvent of the type astaught in the specification, separating the liquid and solid contents,recovering a second lipid rich fraction by evaporation of the solventfrom the liquid contents and recovering the solid contents. Theresultant krill oil extract has also been used in an attempt to decreaselipid profiles in patients with hyperlipidemia. The '808 publicationgives details regarding this krill oil as derived using those generalsteps identified above.

The published article gives further details of how the processed krilloil alone, at 3000 mgs/daily dose is a product that aids in treatingchronic inflammation and arthritic symptoms. The article describes astudy, which had several objectives: a) to evaluate the effect ofNeptune Krill Oil on C-reactive protein (C-RP) on patients with chronicinflammation; and b) to evaluate the effectiveness of the Neptune KrillOil on arthritic symptoms. The method used a randomized, double blind,placebo controlled study protocol. Ninety patients were recruited witheither a confirmed diagnosis of cardiovascular disease and/or rheumatoidarthritis and/or osteoarthritis and with increased levels of CRP (>1.0mg/dl) upon three consecutive weekly blood analysis prior to initiationof oral treatment with krill oil. It is important to note that C-RP is awell known biomarker for risk of cardiovascular disease, therefore inthis trial, since patients with known cardiovascular disease states wenot excluded from the trial the protocol appears to have evaluated theeffects of krill oil on this cardiovascular risk factor while evaluatingthe effects of krill oil supplementation on the pain and discomfortassociated with OA and RH. Group A received the Neptune Krill Oil (300mg daily) and group B received a placebo. C-RP and Western Ontario andMcMaster Universities (WOMAC) osteoarthritis scores were measured atbaseline and days 7, 14 and 30. After seven days of treatment, theNeptune Krill Oil reduced CRP by 19.3% compared to an increase by 15.7%observed in the placebo group (p=0.049). After 14 and 30 days oftreatment, the Neptune Krill Oil further decreased CRP by 29.7% and30.9% respectively (p<0.001). The CRP levels of the placebo groupincreased to 32.1% after 14 days and then decreased to 25.1% at day 30.The between group difference was statistically significant; p=0.004 atday 14 and p=0.008 at day 30. The application of the processed NeptuneKrill Oil showed a significant reduction in all three WOMAC scores.After seven days of treatment, the Neptune Krill Oil reduced pain scoresby 28.9% (p=0.050), reduced stiffness by 20.3% (p=0.001) and reducedfunctional impairment by 22.8% (p=0.008). The results of that studyindicate that the Neptune Krill Oil at a daily dose of about 300 mgsignificantly inhibits inflammation, reduces arthritic symptoms within ashort treatment period of 7 and 14 days and may be effective in reducingthe risk of cardiovascular disease by reduction of C-RP in the patientpopulation employed. It is desirable if further enhanced effects beaccomplished using krill oil and similar compositions, especially withimproving blood lipid profiles and reducing LDL oxidation.

SUMMARY OF THE INVENTION

In accordance with a non-limiting example, even more beneficial andsynergistic effects for improving blood lipid profiles and reducing LOLhave been found when krill oil and/or marine oil is used in combinationwith astaxanthin or an LDL per-oxidation blocker, or when a mixture offish oil derived, choline based, phospholipd bound omega-3 fatty acidmixture including phospholipid bound polyunsaturated EPA and DHA is usedinstead of krill oil.

The composition and method, in accordance with a non-limiting example,improves blood lipid profiles and reduces low density lipoprotein (LDL)oxidation in humans by administering a therapeutic amount of acomposition comprising krill oil in combination with astaxanthin or inanother embodiment, an LDL per-oxidation blocker in an oral dosage form.In one embodiment, the krill oil is derived from Euphasia spp.,comprising Eicosapentaenoic (EPA) and Docosahexaenoic (DNA) fatty acidsin the form of triacylglycerides and phospholipids. The krill oilincludes at least 10% EPA and 5% DHA, of which greater than 50% are inthe form of phospholipids and the 1-4000 mg of krill oil per daily doseis delivered.

In another example, the LDL per-oxidation blacker comprises astaxanthin.In an example, 0.1-12 mg astaxanthin is supplemented to the krill oilper daily dose. The astaxanthin is derived from Haematococcus pluvialisalgae, Pfaffia, krill, or by synthetic routes, in the known free diol,monoester or diester forms. In another example, the compositionadditionally includes a plant based, triacylglycerides bound, n-3(omega-3) fatty acid rich oil derived from flax seed oil, perilla seedoil or chia seed oil wherein the n-3 fatty acid comprises principallyalpha-linolenic acid.

In yet another example, a method and composition is disclosed thatimproves blood lipid profiles and reduces low density lipoprotein (LDL)oxidation in humans by administering a composition comprising atherapeutic amount of a mixture of fish oil derived choline based,phospholipid bound omega-3 fatty acid mixture including phospholipidbound polyunsaturated Eicosapentaenoic (EPA) and Docosahexaenoic (DHA).The fish oil derived, choline based, phospholipid bound fatty acidmixture includes polyunsaturated EPA and DHA and comprisesEicosapentaenoic (EPA) and Docosahexaenoic (DHA) fatty acids in the formof triacylglycerides and phospholipids. The omega choline comprises notless than (n.l.t.) 15 g/100 g of marine phospholipids, n.l.t. 12 g/100 gof DHA, and n.l.t. 7 g/100 g EPA in one example. The omega cholinecomprises n.l.t. 22 g/100 g of Omega-3 and less than 3 g/100 g ofOmega-6. In yet another example, the composition of fish oil derived,choline based, phospholipid bound fatty acid mixture can be enriched inthe phospholipid fraction by the use of supercritical solvent extractionof triacylglycerides from the phospholipid fraction to decrease thetriacylglyceride based fish oil diluents and increase the fish oilderived phospholipids.

Astaxanthin may also be added to the composition fish oil derived,choline based, phospholipid bound fatty acid mixture includingpolyunsaturated EPA and DHA per daily dose. About 0.1-20 mg ofastaxanthin are added to the fish oil derived, choline based,phospholipid bound fatty acid mixture including polyunsaturated EPA andDHA per daily dose. The astaxanthin may be derived from Haematococcuspluvialis algae, Pfaffia, krill, or by synthetic routes, in the freediol, monoester or diester forms.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present invention will now be described more fully hereinafter inwhich preferred embodiments of the invention are described. Thisinvention may, however, be embodied in many different forms and shouldnot be construed as limited to the embodiments set forth herein. Rather,these embodiments are provided so that this disclosure will be thoroughand complete, and will fully convey the scope of the invention to thoseskilled in the art.

In an embodiment using krill oil, the composition includes EPA and DHAfunctionalized as marine phospholipids and acyltriglycerides derivedfrom krill, and in one example, includes esterified astaxanthin in anoral dosage form. It has been found that a new and potentially quiteimportant biomarker for cardiovascular risk is related to the amount ofEPA and DHA found in red blood cells divided by the total fatty acidcontent in red blood cells or the so called “omega-3 index.” Thecompositions, in accordance with a non-limiting example, improve theomega-3 index in man on prolonged administration and therefore arepresumed to lower cardiovascular event risks. Some of these componentsare explained in the following chart:

Components Percentage (%) PHOSPHOLIPIDS >40 PC, PE, PI, PS, SM, CLOMEGA-3 (functionalized on PL) >30 Eicosapentaenoid Acid (EPA)* >17 (15%in one example and 10% in another) Docosahexaenoid Acid (DHA)+ >11 (9%in one example and 5% in another) ANTIOXIDANTS (mg/100 g) Astaxanthin,Vitamin A, Vitamin E  >1.25 *>55% of PL-EPA/Total EPA +>55% ofPL-DHA/Total DHA These amounts can vary depending on application andpersons.

Krill oil can be supplemented with astaxanthin to improve formulatedproduct utility. In one study, 4 mg of astaxanthin per day for two weeksresulted in a 26% reduction of LDL cholesterol oxidation. 4 mg ofastaxanthin for eight weeks resulted in a 21% decrease in C-reactiveprotein scores. 3.6 mg of astaxanthin per day for two weeks demonstratedthat astaxanthin protects LDL cholesterol against induced in vitrooxidation.

Astaxanthin is also known to reduce C-Reactive Protein (C-RP) bloodlevels in vivo. For example, in human subjects with high risk levels ofC-RP three months of astaxanthin treatment resulted in 43% drop in thepatient population's serum C-RP levels a drop which is below theunacceptable cardiovascular event risk level. Astaxanthin is so powerfulthat it has been shown to negate the pro-oxidant activity of Vioxx invitro, a COX-2 inhibitor belonging to the NSAIDS drug class which isknown to cause cellular membrane lipid per-oxidation leading to heartattacks and strokes. For this reason Vioxx was subsequently removed fromthe US market by the FDA. Astaxanthin is also absorbed in vitro by lensepithelial cells where it suppresses UVB induced lipid per-oxidativemediated cell damage at umol/L concentrations. Reduction of C-Reactiveprotein (CRP), reduction of LDL oxidation and an increase in the omega-3index in vivo would presumably all be important positive contributors tocardiovascular health since each are well know biomarkers forcardiovascular health risk. These results have been shown in:

-   1) Lee et al., Molecules and Cells, 16(1):97-105; 2003;-   2) Ohgami et al., Investigative Ophthalmology and Visual Science    44(6):2694-2701, 2003;-   3) Spiller et al., J. of the Amer. College of Nutrition, 21(5):    October 2002; and-   4) Harris, Pharmacol. Res. 2007 March; 55(3) 217-223.

A preferred composition in one embodiment includes 300-500 mg of krilloil and 2 mg astaxanthin.

As noted before, krill oil is typically produced from Antarctic krill(Euphausia superba), which is a zooplankton (base of food chain). It isone of the most abundant marine biomass of about 500 million tonsaccording to some estimates. Antarctic krill breeds in the pureuncontaminated deep sea waters. It is a non-exploited marine biomass andthe catch per year is less than or equal to about 0.02% according tosome estimates.

It is believed that Krill oil based phospholipid bound EPA and DHAuptake into cellular membranes is far more efficient thantriacylglyercide bound EPA and DHA since liver conversion oftriacylglycerides is itself inefficient and because phospholipid boundEPA and DHA can be transported into the blood stream via the lympathicsystem, thus, avoiding liver breakdown. In addition, krill oilconsumption does not produce the burp-back observed with fish oil basedproducts. Because of this burp-back feature of fish oils, it has beenfound that approximately 50% of all consumers who try fish oil never buyit again.

Astaxanthin has an excellent safety record. A conducted study obtainedthe results as follows:

Oral LD 50: 600 mg/kg (rats); NOAEL: 465 mg/kg (rats); or SerumPharmacokinetics: Stewart et al. 2008 1) T_(1/2): 16 hours; 2) T_(max):8 hours; 3) C_(max): 65 μg/L.

At eight weeks of supplementation at 6 mg per day, there was no negativeeffect in healthy adults. Spiller et al. 2003.

In accordance with one non-limiting example, astaxanthin has three primesources. 3 mg astaxanthin per 240 g serving of non-farmed raised salmonor a 1% to 12% astaxanthin oleoresin or 1.5-2.5% beadlet derived frommicroalgae. Literature references pertinent to the above discussion canbe found in Lee et al., Molecules and Cells 16(1): 97-105, 2003; Ohgamiet al., Investigative Ophthalmology and Visual Science 44(6): 2694-2701,2003; Spiller et al., J. of the American College of Nutrition 21(5):October 2002; and Fry et al., University of Memphis, Human PerformanceLaboratories, 2001 and 2004, Reports 1 and 2.

Although many beneficial and synergistic effects are now being reportedherein have been observed when krill oil is used in combination withother active ingredients, and more specifically in one example, krilloil in combination with astaxanthin. It should be understood thatdifferent proportions of ingredients and percentages in compositions canbe used depending on end use applications and other environmental andphysiological factors when treating a patient condition.

The krill oil in one example is derived from Euphasia spp., comprisingEicosapentaenoic (EPA) and Docosahexaenoic (DHA) fatty acids in the formof triacylglycerides and phospholipids, although not less than 1% EPAand 5% DHA has been found advantageous. In another example, the krilloil includes at least 15% EPA and 9% DHA, of which not less than 45% arein the form of phospholipids, and in one example, greater than 50%. Thecomposition can be delivered advantageously for therapeutic results with1-4000 mg of krill oil delivered per daily dose. In another example,0.1-50 mg astaxanthin are supplemented to the krill oil per daily dose,and in one example, 0.1-12 mg of astaxanthin.

The astaxanthin is preferably derived from Haematococcus pluvialisalgae, Pfaffia, krill, or by synthetic routes, in the known free diol,monoester or diester form, and in one example, at a daily dose of 0.5-Bmg.

The composition may also include an n-3 (omega-3) fatty acid rich oilderived from fish oil, algae oil, flax seed oil, or chia seed oil whenthe n-3 fatty acid comprises alpha-linolenic, stearidonic,eicosapentaenoic or docosapentaenoic acid. The composition may includenaturally-derived and synthetic antioxidants that are added to retarddegradation of fatty acids and astaxanthin.

Details of a type of CO2 extraction and processing technology (assupercritical CO2 extraction) and peroxidation blocker technology thatcan be used are disclosed in commonly assigned U.S. Patent PublicationNos. 2009/0181127; 2009/0181114; and 2009/0258081, the disclosures whichare hereby incorporated by reference in their entirety.

As noted before, there are beneficial aspects of using krill oil insynergistic combination with other ingredients. It has been determinedthat a fish oil derived, choline based, phospholipid bound omega-3 fattyacid mixture including phospholipid bound polyunsaturated EPA and DHA isalso advantageous for improving blood lipid profiles and reducing LDLeither alone or admixed with other ingredients, for example, an LDLper-oxidation blocker. One commercially available example of a mixtureof fish oil derived, choline based, phospholipid bound fatty acidmixture including polyunsaturated EPA and DHA is Omega Choline 1520F asa phospholipid, omega-3 preparation, which is derived from natural fishoil and sold by Enzymotec Ltd. One example of such composition isdescribed below:

Ingredients (g/100 g): Pure Marine Phospholipids n.l.t. 15 DHA* n.l.t.12 EPA** n.l.t. 7 Omega-3 n.l.t. 22 Omega-6 <3 Analytical Data: Peroxidevalue (meq/Kg) n.m.t. 5 Loss on Drying (g/100 g) n.m.t. 2 PhysicalProperties: Consistency Viscous Liquid *Docosahexaenoic acid**Eicosapenteanoic acid

In accordance with a non-limiting example, the method improves bloodlipid profiles and either alone or in combination with addedastaxanthin, such as a per-oxidation blocker, and reduces LDL oxidationin a patient by administering a therapeutic amount of a compositionincluding a mixture of fish oil derived, choline based, phospholipidbound omega-3 fatty acid mixture including phospholipid boundpolyunsaturated EPA and DHA either alone or admixed with an LDLper-oxidation blocker such as astaxanthin. In one example, thecomposition is supplemented in combination with astaxanthin in an oraldosage form. The mixture of fish oil derived, choline based,phospholipid bound fatty acid mixture including polyunsaturated EPA andDHA in one example comprises Eicosapentaenoic (EPA) and Docosahexaenoic(DHA) fatty acids in the form of triacylglycerides and phospholipids. Inanother example, the omega choline includes at least 7% EPA and 12% DHA,of which not less than 15% are in the form of phospholipids. Thecomposition can be delivered advantageously for therapeutic results with1-4000 mg of a mixture of fish oil and fish oil derived, choline based,phospholipid bound fatty acid mixture including polyunsaturated EPA andDHA delivered per daily dose. In another example, 0.1-20 mg astaxanthinare supplemented to the Omega Choline per daily dose.

It should be understood that an instant formulation can be used for LDLreduction using only a mixture of fish oil derived, choline based,phospholipid bound fatty acid mixture including polyunsaturated EPA andDHA. It is also possible to use a mixture of fish oil derived, cholinebased, phospholipid bound omega-3 fatty acid mixture (includingpolyunsaturated EPA and DHA) mixed with astaxanthin. It should also beunderstood that an enriched version of a mixture of fish oil derived,choline based, phospholipid bound fatty acid mixture includingpolyunsaturated EPA and DHA can be used wherein the fraction of addedfish oil diluents has been decreased and the proportion of fish oilderived phospholipids has been increased. This can be accomplished byusing supercritical CO2 and/or solvent extractions for selective removalof triacylglycerides from phospholipids. The composition may alsoinclude a natural or synthetic cyclooxygenase-1 or -2 inhibitorcomprising for example aspirin, acetaminophen, steroids, prednisone, orNSAIDs. The composition may also include a gamma-linoleic acid rich oilcomprising Borage (Borago officinalis L.) or Safflower (Carthamustinctorius L.), which delivers a metabolic precursor to PGE₁ synthesis.

The composition may also include an n-3 (omega-3) fatty acid rich oilderived from fish oil, algae oil, flax seed oil, chia seed oil orperilla seed oil wherein the n-3 fatty acid source comprisesalpha-linolenic, stearidonic, eicosapentaenoic or docosapentaenoic acid.The composition may include naturally-derived and synthetic antioxidantsthat are added to retard degradation of fatty acids such as tocopherols,tocotrienols, carnosic acid or Carnosol and/or astaxanthin.

Many modifications and other embodiments of the invention will come tothe mind of one skilled in the art having the benefit of the teachingspresented in the foregoing descriptions and the associated drawings.Therefore, it is understood that the invention is not to be limited tothe specific embodiments disclosed, and that modifications andembodiments are intended to be included within the scope of the appendedclaims.

1-8. (canceled)
 9. A composition to treat low density lipoprotein (LDL)oxidation in humans, wherein the composition includes a therapeuticamount of krill oil in combination with astaxanthin in an oral dosageform.
 10. The composition according to claim 9, wherein the krill oil isderived from Euphasia spp., comprising Eicosapentaenoic (EPA) andDocosahexaenoic (DHA) fatty acids in the form of triacylglycerides andphospholipids.
 11. The composition according to claim 9, wherein thekrill oil includes at least 10% EPA and 5% DHA, of which greater than40% are in the form of phospholipids.
 12. The composition according toclaim 9, wherein the composition is formulated to deliver 1-4000 mg ofkrill oil.
 13. (canceled)
 14. The composition according to claim 13,wherein the composition includes 0.1-12 mg astaxanthin supplemented tothe krill oil.
 15. The composition according to claim 13, wherein theastaxanthin is derived from Haematococcus pluvialis algae, Pfaffia,krill, or by synthetic routes, in the known free diol, monoester ordiester form.
 16. The composition according to claim 9, and furthercomprising an n-3 (omega-3) fatty acid rich oil derived from flax seedoil, perilla seed oil or chia seed oil wherein the n-3 fatty acidcomprises alpha-linolenic acid. 17-23. (canceled)
 24. A compositionformulated in a therapeutic amount to treat low density lipoprotein(LDL) oxidation in humans, wherein the composition comprises a mixtureof fish oil derived, choline based, phospholipid bound omega-3 fattyacid mixture including phospholipid bound polyunsaturatedEicosapentaenoic (EPA) and Docosahexaenoic (DHA) either alone or incombination with a per-oxidation blocker.
 25. The composition accordingto claim 24, wherein the Omega Choline comprises Eicosapentaenoic (EPA)and Docosahexaenoic (DHA) fatty acids bound in the form of bothtriacylglycerides and phospholipids and collectively derived from fishoil.
 26. The composition according to claim 24, wherein the mixture offish oil derived, choline based, phospholipid bound omega-3 fatty acidmixture including phospholipid bound polyunsaturated EPA and DHAcomprises not less than (n.l.t.) 15 g/100 g of phospholipids, n.l.t. 12g/100 g of DHA, and n.l.t. 7 g/100 g EPA.
 27. The composition accordingto claim 26, wherein the mixture of fish oil derived, choline based,phospholipid bound omega-3 fatty acid mixture including phospholipidbound polyunsaturated EPA and DHA comprises n.l.t. 22 g/100 g ofOmega-3.
 28. The composition according to claim 24, wherein the mixtureof fish oil derived, choline based, phospholipid bound omega-3 fattyacid mixture wherein the phospholipid bound polyunsaturated EPA and DHAincludes a supercritical solvent extracted triacylglycerides bound fattyacid fraction from the remaining phospholipids to decrease thetriacylglycerides bound fatty acid fish oil fraction and therebyincreasing the fish oil derived phospholipids.
 29. The compositionaccording to claim 24, wherein the mixture of fish oil derived, cholinebased, phospholipid bound omega-3 fatty acid mixture includingphospholipid bound polyunsaturated EPA and DHA includes at least 7% EPAand 12% DHA, of which not less than 15% are in the form ofphospholipids.
 30. (canceled)
 31. The composition according to claim 24,wherein the composition includes an n-3 (omega-3) fatty acid rich fishoil, algae oil, flax seed oil, perilla seed oil or chia seed oil and then-3 fatty acid comprises alpha-linolenic, stearidonic, eicosapentaenoicor docosapentaenoic acid.
 32. The composition according to claim 24, andfurther comprising astaxanthin.
 33. The composition according to claim32, and further comprising 0.1-12 mg astaxanthin supplemented to thekrill oil.
 34. The composition according to claim 32, wherein theastaxanthin is derived from Haematococcus pluvialis algae, Pfaffia,krill, or by synthetic routes, in the known free diol, monoester ordiester form.
 35. A composition formulated in a therapeutic amount totreat low density lipoprotein (LDL) oxidation in humans, wherein thecomposition comprises a mixture of fish oil derived, choline based,phospholipid bound omega-3 fatty acid mixture including phospholipidbound polyunsaturated Eicosapentaenoic (EPA) and Docosahexaenoic (DHA)acids.
 36. The composition according to claim 35, wherein the OmegaCholine comprises Eicosapentaenoic (EPA) and Docosahexaenoic (DHA) fattyacids bound in triacylglycerides and phospholipids and derived from fishoil.
 37. The composition according to claim 35, wherein the mixture offish oil derived, choline based, phospholipid bound omega-3 fatty acidmixture including phospholipid bound polyunsaturated EPA and DHAcomprises not less than (n.l.t.) 15 g/100 g of phospholipids, n.l.t. 12g/100 g of DHA, and n.l.t. 7 g/100 g EPA.
 38. The composition accordingto claim 35, wherein the mixture of fish oil derived, choline based,phospholipid bound omega-3 fatty acid mixture including phospholipidbound polyunsaturated EPA and DHA comprises n.l.t. 22 g/100 g ofOmega-3.
 39. The composition according to claim 35, wherein the mixtureof fish oil derived, choline based, phospholipid bound omega-3 fattyacid mixture wherein the phospholipid bound polyunsaturated EPA and DHAincludes a supercritical solvent extracted triacylglycerides bound fattyacid fraction from the remaining phospholipids to decrease thetriacylglycerides bound fatty acid fish oil fraction and therebyincreasing the fish oil derived phospholipids.
 40. The compositionaccording to claim 35, wherein the mixture of fish oil derived, cholinebased, phospholipid bound omega-3 fatty acid mixture includingphospholipid bound polyunsaturated EPA and DHA includes at least 7% EPAand 12% DHA, of which not less than 15% are in the form ofphospholipids.
 41. (canceled)
 42. The composition according to claim 35,and further comprising astaxanthin.